Target-induced clustering activates Trim-Away of pathogens and proteins

Zeng, Jingwei; Santos, Ana Filipa; Mukadam, Aamir S.; Osswald, Mariana; Jacques, D.A.; Dickson, Claire F.; McLaughlin, Stephen H.; Johnson, C.M.; Kiss, Leo; Lupták, Jakub; Renner, Nadine; Vaysburd, Marina; McEwan, William A.; Morais-de-Sá, Eurico; Clift, Dean; James, Leo C.

doi:10.1038/s41594-021-00560-2

Cited by 45 publications

(91 citation statements)

References 63 publications

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“…Studies have shown dimerisation of the catalytic RING domains to be necessary for ubiquitinating activity in most TRIM proteins studied so far (48). This holds true for TRIM21, with a recent study demonstrating that dimerisation at TRIM21's coiled-coil directly enabled catalytic activity (49). Mutation of residues at the antiparallel coiled-coil dimer interface was sufficient to inhibit auto-ubiquitination of the RING domain, following infection of HEK293T cells with antibody-coated adenovirus.…”

Section: Coiled-coilmentioning

confidence: 93%

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TRIM21/Ro52 - Roles in Innate Immunity and Autoimmune Disease

2021

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TRIM21 (Ro52/SSA1) is an E3 ubiquitin ligase with key roles in immune host defence, signal transduction, and possibly cell cycle regulation. It is also an autoantibody target in Sjögren’s syndrome, systemic lupus erythematosus, and other rheumatic autoimmune diseases. Here, we summarise the structure and function of this enzyme, its roles in innate immunity, adaptive immunity and cellular homeostasis, the pathogenesis of autoimmunity against TRIM21, and the potential impacts of autoantibodies to this intracellular protein.

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Section: Coiled-coilmentioning

confidence: 93%

“…Furthermore, forced RING domain dimerisation ( via RING-linker-RING constructs) increased ubiquitin discharge activity. A proposed model of “clustering-induced activation” suggests dimerisation of RING domains makes E2-ubiquitin engagement with TRIM21 and subsequent polyubiquitin discharge more energetically favourable ( 49 ).…”

Section: Trim21 Structurementioning

confidence: 99%

TRIM21/Ro52 - Roles in Innate Immunity and Autoimmune Disease

2021

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“…An OptoTrim-Away construct, which contains TRIM21 fused with CRY2 and an anti-GFP nanobody vhhGFP4, has been generated to demonstrate light-inducible degradation of GFP-tagged protein ( FIGURE 10 C ). Given that the RING domain alone is sufficient to trigger clustering and proteasomal degradation, a minimal version of OptoTrim-Away was created to cause target degradation to a similar level of full-length TRIM21 ( 349 ). Red or far-red light-inducible degradation systems are yet to be developed to enable paralleled degradation of multiple endogenous targets in the same cells.…”

Section: Application Of Optogenetics In Physiological Processesmentioning

confidence: 99%

Optophysiology: Illuminating cell physiology with optogenetics

Tan

Huang

et al. 2022

Physiological Reviews

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Optogenetics combines light and genetics to enable precise control of living cells, tissues, and organisms with tailored functions. Optogenetics has the advantages of noninvasiveness, rapid responsiveness, tunable reversibility, and superior spatiotemporal resolution. Following the initial discovery of microbial opsins as light-actuated ion channels, a plethora of naturally occurring or engineered photoreceptors or photosensitive domains that respond to light at varying wavelengths has ushered in the next chapter of optogenetics. Through protein engineering and synthetic biology approaches, genetically-encoded photoswitches can be modularly engineered into protein scaffolds or host cells to control a myriad of biological processes, as well as to enable behavioral control and disease intervention in vivo. Here, we summarize these optogenetic tools on the basis of their fundamental photochemical properties to better inform the chemical basis and design principles. We also highlight exemplary applications of opsin-free optogenetics in dissecting cellular physiology (designated "optophysiology"), and describe the current progress, as well as future trends, in wireless optogenetics, which enables remote interrogation of physiological processes with minimal invasiveness. This review is anticipated to spark novel thoughts on engineering next-generation optogenetic tools and devices that promise to accelerate both basic and translational studies.

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“…TRIM21 recognizes antibody-bound pathogens through a high-affinity interaction with the Fc domain of the antibody and tags the antibody-bound pathogens for ubiquitination and degradation. Clift et al repurposed TRIM21 to establish a method called Trim-Away that targets endogenous proteins for proteasomal degradation using endogenous or exogenous TRIM21 and antibodies that bind to the POI (Figure 2b) [30,31]. Notably, this technique enables the selective degradation of post-translationally modified protein and splice or mutant protein variants, by using a specific antibody, while preserving the unmodified/wild-type protein.…”

Section: Antibody-based Approachesmentioning

confidence: 99%

Strategies for Post-Translational Control of Protein Expression and Their Applications

Utsugi

Miyamae

2021

Applied Sciences

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Proteins are fundamental biomolecules of living cells, and their expression levels depend on the balance between the synthesis and degradation. Researchers often aim to control protein expression levels for the investigation of protein function and its relationship with physiological phenomena. The genetic manipulation of the target protein using CRISPR/Cas9, Cre/loxP, tetracyclin system, and RNA interference, are widely used for the regulation of proteins at the DNA, transcriptional, or mRNA level. However, the significant time delay in controlling protein levels is a limitation of these techniques; the knockout or knockdown effects cannot be observed until the previously transcribed and synthesized protein is degraded. Recently, researchers have developed various types of molecular tools for the regulation of protein expression at the post-translational level, which rely on harnessing cellular proteolytic machinery including ubiquitin–proteasome pathway, autophagy-lysosome pathway, and endocytosis. The post-translational control of protein expression using small molecules, antibodies, and light can offer significant advantages regarding speed, tunability, and reversibility. These technologies are expected to be applied to pharmacotherapy and cell therapy, as well as research tools for fundamental biological studies. Here, we review the established and recently developed technologies, provide an update on their applications, and anticipate potential future directions.

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Target-induced clustering activates Trim-Away of pathogens and proteins

Cited by 45 publications

References 63 publications

TRIM21/Ro52 - Roles in Innate Immunity and Autoimmune Disease

TRIM21/Ro52 - Roles in Innate Immunity and Autoimmune Disease

Optophysiology: Illuminating cell physiology with optogenetics

Strategies for Post-Translational Control of Protein Expression and Their Applications

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